CN114223729B - Application of bifidobacterium longum subspecies infantis and breast milk oligosaccharide composition in formula milk powder - Google Patents

Application of bifidobacterium longum subspecies infantis and breast milk oligosaccharide composition in formula milk powder Download PDF

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CN114223729B
CN114223729B CN202111447747.8A CN202111447747A CN114223729B CN 114223729 B CN114223729 B CN 114223729B CN 202111447747 A CN202111447747 A CN 202111447747A CN 114223729 B CN114223729 B CN 114223729B
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formula
bifidobacterium longum
powder
breast milk
milk
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CN114223729A (en
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郝威
刘彪
李威
孔小宇
肖竞舟
吉塞拉·阿德里安娜·怀斯
卡洛琳·安妮卡·凡·鲁-鲍曼
盖瑞特·施密特
王雯丹
司徒文佑
刘伟贤
洪维鍊
王燕霞
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Inner Mongolia Yili Industrial Group Co Ltd
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Inner Mongolia Yili Industrial Group Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/06—Mixtures of whey with milk products or milk components
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/02—Whey; Whey preparations containing, or treated with, microorganisms or enzymes
    • A23C21/026—Whey; Whey preparations containing, or treated with, microorganisms or enzymes containing, or treated only with, lactic acid producing bacteria, bifidobacteria or propionic acid bacteria
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/04—Whey; Whey preparations containing non-milk components as source of fats or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/08—Whey; Whey preparations containing other organic additives, e.g. vegetable or animal products
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/10—Whey; Whey preparations containing inorganic additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/531—Lactis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/533—Longum

Abstract

The invention provides an application of a bifidobacterium longum subspecies infantis and breast milk oligosaccharide composition in formula milk powder. In particular, the invention provides the application of a composition of bifidobacterium longum subspecies infantis and breast milk oligosaccharide in preparing the formula milk powder which can effectively improve the resistance of organisms to staphylococcus aureus infection and can improve the innate immunity and/or anti-aging efficacy of organisms, wherein each 100g of the formula milk powder contains 1X 10 of bifidobacterium longum subspecies infantis 3 CFU～1×10 12 CFU, 0.1 g-10 g of breast milk oligosaccharide.

Description

Application of bifidobacterium longum subspecies infantis and breast milk oligosaccharide composition in formula milk powder
Technical Field
The invention relates to application of a bifidobacterium longum subspecies infantis and breast milk oligosaccharide composition in preparation of formula milk powder.
Background
In the last thousand years, medical literature has described that non-breast-fed infants have a higher disease rate and mortality rate than breast-fed infants. The breast milk not only provides needed nutrition for infants, but also provides guarantee for intestinal development and immunity improvement of infants due to active ingredients in the breast milk. Breast-fed infants have a higher relative abundance of beneficial bacteria, particularly bifidobacteria and lactic acid bacteria, in the intestinal flora than formula-fed infants.
The breast milk is transmitted by flora, and active ingredients such as breast milk oligosaccharide and cytokines in the breast milk are added, so that a healthy intestinal flora is established for the newborn. Infants ingest 10 a day by breast milk 7 -10 8 Bacteria, including lactic acid bacteria and bifidobacteria. The bacteria are directly transmitted to the infant through breast milk, part of the bacteria can colonize the intestinal tracts of the infant, and the establishment of intestinal flora in early life is promoted. The establishment of the intestinal flora of infants has a short-term, even lifetime, impact on the development of their intestinal tract, as well as on the health and immune system.
Breast milk oligosaccharides (Human Milk Oligosaccharides, abbreviated HMOs) are the third largest population of milk compounds present in human milk. It plays an important role in early infant development. To date, 130 different structures have been identified, but the functional impact is unknown. Most oligosaccharides in milk cannot be digested and therefore HMO can act as a prebiotic. Furthermore, HMO shows its importance for healthy development in infants in terms of maturation of the immune system and its role as immunomodulator. Currently, HMOs have been added to infant formulas.
At present, in the fields of infant formulas, complementary foods and nutritional supplements, solutions for relieving intestinal discomfort of infants and improving the capability of self-defense against pathogenic bacteria such as staphylococcus aureus infection are needed. Meanwhile, in the fields of children, teenagers and adults over 3 years old, solutions for relieving intestinal discomfort and improving the ability of themselves to fight against pathogenic bacteria such as staphylococcus aureus are also needed.
Caenorhabditis elegans (c.elegans) as a model organism has a good application prospect in preclinical research and evaluation. It has short life cycle (21 days), high replicability and reproducibility, simple operation, transparency and easy cultivation. Its genome has been fully sequenced and one quarter of the genes are homologous to the human genome. The nematode organism with the genetic mutation generated by editing the nematode gene can be used as an experimental means for genetic analysis. Nematodes are not currently considered an animal in european legislation. It is widely used as an in vitro assay such as transcriptomics, proteomics, metabolomics, etc. As a model organism, it is also often used as the first step in raw material evaluation, and nematodes are often used as a high throughput means to screen test raw materials for some properties prior to design of efficacy raw materials, in vitro enzyme or cell experiments, mouse models and clinical trials.
Staphylococcus aureus (Staphylococcus aureus, s.aureus) is a gram positive bacterium belonging to the genus staphylococcus and is a common food-borne pathogenic microorganism. It is widely found in natural environments, under appropriate conditions, can produce enterotoxins and can cause food poisoning, causing excessive food-borne microbial food poisoning events caused by staphylococcus aureus. After the human body is infected by staphylococcus aureus, common food poisoning symptoms such as nausea, vomit, dizziness and the like can occur, and symptoms such as enteritis, pneumonia, skin infection, wound infection ulcer, meningitis and the like can occur.
Disclosure of Invention
The inventors have found in research that some bifidobacterium longum subspecies infantis in combination with breast milk oligosaccharides enhance the ability of an organism to resist staphylococcus aureus infection, thereby providing the following invention:
in one aspect, the invention provides the use of a composition of bifidobacterium longum subspecies infantis and breast milk oligosaccharides for the preparation of a formula having an effect of effectively improving the resistance of an organism to staphylococcus aureus infection, improving the innate immunity and/or anti-aging efficacy of the organism, wherein each 100g of the formula contains 1X 10 bifidobacterium longum subspecies infantis 3 CFU～1×10 12 CFU/100g, 0.1 g-10 g of breast milk oligosaccharide.
On the other hand, the invention also provides a formula milk powder which can effectively improve the resistance of organisms to staphylococcus aureus infection and can improve the innate immunity and/or anti-aging effect of organisms, and the formula milk powder is prepared by adding bifidobacterium longum subspecies and breast milk oligosaccharide into the formula milk powder.
According to a specific embodiment of the present invention, in the formula of the present invention, the bifidobacterium longum subspecies infancy comprises bifidobacterium longum subspecies infancy (Bifidobacterium longum subsp.infantis) YLGB-1496 strain with a preservation number of CGMCC No. 21109. The bifidobacterium longum subspecies infantis strain with the preservation number of CGMCC No.21109 is preserved in China general microbiological culture Collection center (CGMCC) at the 12 th month 05 of 2020, and the preservation unit address is: the national institute of microbiology, national academy of sciences, china, collection date: 12 months 05 days 2020; preservation number: CGMCC No.21109; classification naming: bifidobacterium longum subspecies infantis (Bifidobacterium longum subsp.infantis). The strain is also referred to herein as YLGB-1496 or GB-1496.
According to the specific embodiment of the invention, the total protein content in the formula milk powder is 10 g-20 g/100g, and the proportion of whey protein in the total protein is 40% -70%; the fat content is 15 g-30 g/100g; the carbohydrate content is 50 g-70 g/100g.
According to the specific embodiment of the invention, the formula milk powder also contains linoleic acid and alpha-linolenic acid, wherein the linoleic acid content is 2700-4500 mg/100g, and the alpha-linolenic acid content is 270-450 mg/100g.
According to the specific embodiment of the invention, the formula powder can be infant formula powder (infant formula food) or non-infant formula powder (such as children formula powder and adult formula powder).
According to a specific embodiment of the present invention, in the formula of the present invention, the raw materials for providing total protein include: raw milk (cattle/sheep), whole milk powder (cattle/sheep), skimmed milk powder (cattle/sheep), whey protein powder (cattle/sheep), desalted whey powder (cattle/sheep), and beta-casein. Specifically, based on 1000 parts by weight of the formula milk powder, the raw materials comprise: raw milk (cattle/sheep) 0-3500 parts, whole milk powder 0-260 parts (cattle/sheep), whey protein powder (cattle/sheep) 0-120 parts, skim milk powder (cattle/sheep) 0-400 parts, and/or desalted whey powder (cattle/sheep) 0-600 parts, with the addition of beta-casein being 0-40 parts.
According to a specific embodiment of the invention, the raw materials for providing fat in the formula milk powder comprise one or more of the following raw materials in parts by weight: 0-50 parts of corn oil, 0-65 parts of soybean oil, 0-100 parts of sunflower seed oil, 0-240 parts of 1, 3-dioleoyl-2-palmitic acid triglyceride, 0-30 parts of coconut oil, 0-85 parts of canola oil and 0-20 parts of linseed oil.
According to a specific embodiment of the present invention, the starting materials for providing carbohydrates in the formula according to the present invention comprise: lactose 0-500 parts, and/or the above protein material containing lactose.
According to a specific embodiment of the present invention, the formula of the present invention may comprise, in addition to the above components, conventional components of the formula, for example, one or more of nutrients, dietary fibers, inositol, taurine, l-carnitine, docosahexaenoic acid, eicosatetraenoic acid, lutein, nucleotides, lactoferrin. Preferably comprising a reconstituted nutrient package. In some preferred embodiments, the formula includes 0-20 parts of nutrients including vitamins and minerals, orOne step of selectivity comprises one or more of dietary fiber (fructo-oligosaccharide, galacto-oligosaccharide, fructo-oligosaccharide, etc.), inositol, taurine, L-carnitine, docosahexaenoic acid, eicosatetraenoic acid, lutein, nucleotide, lactoferrin. The compound nutrient mainly comprises the following components: and (3) compounding vitamins: vitamin A, vitamin D, vitamin E, vitamin K 1 Vitamin B 1 Vitamin B 2 Vitamin B 6 Vitamin B 12 Niacin, folic acid, pantothenic acid, vitamin C, choline, biotin; compounding minerals: sodium, potassium, calcium, phosphorus, copper, iron, zinc, manganese, iodine, selenium, magnesium, potassium, and derivatives thereof. The sources, the contents and the usage amount of each vitamin and mineral substance should meet the national standard and related regulations of infant formula food.
According to a specific embodiment of the present invention, the present invention is applied in a composition of bifidobacterium longum subspecies infancy and breast milk oligosaccharides comprising one or more of 2'-fucosyllactose, 3' -fucosyllactose, lactose-N-tetraose, 3'-sialyllactose, 6' -sialyllactose for the preparation of a formula.
2'-fucosyllactose (2' -FL) is a trisaccharide structure formed by fucose and lactose, and is a representative substance of fucosyl oligosaccharides. The commercial material is typically prepared by microbial fermentation and has the same structure as the 2' -fucosyllactose found in human milk.
3-fucosyllactose (3-FL) is a trisaccharide structure formed by fucose and lactose, and is an isomer with 2' -fucosyllactose, and is a representative substance of fucosyl oligosaccharides. The commercial material is typically prepared by microbial fermentation and has the same structure as 3-fucosyllactose found in human milk.
lactose-N-tetraose (LNT), a representative substance of oligosaccharides based on a core sugar chain and free of fucosyl or sialyl groups, is generally prepared by microbial fermentation and has the same structure as lactose-N-tetraose found in human milk.
3' -sialyllactose (3 ' -SL), which is a trisaccharide structure formed by sialic acid and lactose, is an isomer with 6' -sialyllactose, and is a representative substance of sialyl oligosaccharides. The commercial material is usually prepared by microbial fermentation and has the same structure as 3' -sialyllactose found in human milk.
6' -sialyllactose (6 ' -SL), which is a trisaccharide structure formed by sialic acid and lactose, is an isomer with 3' -sialyllactose, and is a representative substance of sialyl oligosaccharides. The commercial material is usually prepared by microbial fermentation and has the same structure as 6' -sialyllactose found in human milk.
According to a specific embodiment of the invention, the present invention is applied in a composition of bifidobacterium longum subspecies infancy and breast milk oligosaccharides comprising 2' -fucosyllactose for the preparation of a formula. The content of 2' -fucosyllactose in the breast milk oligosaccharide may be 80% or more, for example 85% or more, 90% or more, 95% or more, or 100%.
According to a specific embodiment of the present invention, the present invention is applied in a composition of bifidobacterium longum subspecies infancy and breast milk oligosaccharides in the preparation of a formula, said breast milk oligosaccharides being a combination consisting of at least two, at least three or at least four of 2'-fucosyllactose (2' -FL), 3-fucosyllactose (3-FL), lactose-N-tetraose (LNT), 3'-sialyllactose (3' -SL) and 6'-sialyllactose (6' -SL).
According to a specific embodiment of the present invention, the present invention is applied in the composition of Bifidobacterium longum subspecies infantis and human milk oligosaccharides in a ratio of 1X 10 in the preparation of a formula 3 CFU～1×10 12 CFU:0.1g to 10g, preferably 1X 10g 6 CFU～1×10 10 CFU:1 g-10 g. In some more specific embodiments of the invention, the ratio of bifidobacterium longum subspecies infantis to breast milk oligosaccharides is 1X 10 8 CFU:0.08 g-0.3 g. The bifidobacterium longum subspecies infantis and breast milk oligomerizationThe ratio of sugar is calculated by the amount of the two in the same composition.
According to a specific embodiment of the invention, the use of the composition of the invention for increasing the resistance of an organism to staphylococcus aureus infection comprises: improving the ability of an individual to prevent staphylococcus aureus infection, reducing the ability of an individual to be infected with staphylococcus aureus infection, and/or alleviating various symptoms caused by staphylococcus aureus infection, such as food poisoning (including nausea, vomiting, dizziness, etc.), enteritis, pneumonia, skin infection, wound ulcer, or meningitis, etc.
According to a specific embodiment of the invention, the organism comprises an animal or a human in the use of the composition of the invention.
The formula milk powder provided by the invention has the effects of effectively improving the resistance of organisms to staphylococcus aureus infection, improving the innate immunity of organisms and/or resisting aging due to the composition of the bifidobacterium longum subspecies and the breast milk oligosaccharide.
In another aspect, the invention also provides a method for preparing the formula milk powder, which comprises the step of mixing the bifidobacterium longum subspecies and the breast milk oligosaccharide composition with other raw materials in the formula by adopting a wet method or a dry-wet compound production process to prepare the breast milk formula milk powder. The preparation process mainly comprises the following steps: proportioning, homogenizing, concentrating, sterilizing, spray drying and dry mixing to obtain the finished product. In the present invention, the bifidobacterium longum subspecies may be mixed together at the time of dosing (in inactivated form in the infant formula), preferably during post-mixing (in live form in the infant formula). In the invention, the breast milk oligosaccharide can be mixed together during the mixing process, and can also be added during the post-mixing process.
In summary, the composition comprising bifidobacterium longum subspecies infantis and breast milk oligosaccharides has the advantages of resisting aging, improving the innate immunity of organisms, effectively improving the resistance of the organisms to staphylococcus aureus infection, resisting infection and improving immunity when applied to formula milk powder, and has wide application prospect.
Drawings
FIG. 1 shows the effect of a 15mg/mL composition of breast milk oligosaccharide 2' -FL and probiotic YLGB-1496 on survival of C.elegans infected with Staphylococcus aureus.
Microbial preservation for patent procedures:
bifidobacterium infantis YLGB-1496 (or GB-1496) strain:
preservation date: 12 months 05 days 2020;
preservation unit: china general microbiological culture Collection center (CGMCC);
deposit unit address: the institute of microbiology, national academy of sciences, north chen xi lu 1,3, the region of the morning sun in beijing;
preservation number: CGMCC No.21109;
classification naming: bifidobacterium longum subspecies infantis (Bifidobacterium longum subsp.infantis).
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which are derived by a person skilled in the art based on the embodiments of the invention, fall within the scope of protection of the invention.
Unless specifically defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the relevant art.
The operating conditions not specified in detail in the examples were carried out according to the usual procedures in the art.
The probiotics used in the invention is bifidobacterium longum subspecies infancy YLGB-1496 (the source is Chinese healthy mother breast milk, which is preserved in China general microbiological culture Collection center (CGMCC) of China Commission for culture Collection of microorganisms (China general microbiological culture Collection center) at 12 th month and 05 th month of the year of preservation: 12 th month of the institute of microbiology, china academy of sciences of China, the preserving date: CGMCC No.21109; and the classification name: bifidobacterium longum subspecies infancy (Bifidobacterium longum subsp.inffantis)).
The HMO feedstock used in the present invention is derived from Jennewein.
Example 1, infant formula containing probiotic and prebiotic composition (preparation 1000 kg):
1000 kg of raw milk, 320 kg of lactose, 25 kg of whey protein powder WPC80%, 175 kg of desalted whey powder D90 kg, 40 kg of corn oil, 50 kg of soybean oil, 140 kg of OPO structural fat, 27 kg of alpha-whey protein powder, 9 kg of beta-casein powder, 1 kg of anhydrous butter, 17 kg of fructo-oligosaccharide powder, 40 kg of galacto-oligosaccharide syrup, 2 kg of breast milk oligosaccharide (2' -FL), 17 kg of compound nutrient, 12 kg of DHA, 22 kg of ARA, 0.1 kg of bifidobacterium longum subspecies YLGB-1496 and 0.1 kg of bifidobacterium lactis.
Wherein the compound nutrients comprise about 2.5 kg of compound vitamin nutrition package, about 0.75 kg of choline chloride nutrition package, about 6 kg of calcium powder nutrition package, about 1 kg of mineral nutrition package, about 1.5 kg of magnesium chloride nutrition package and about 2 kg of potassium chloride nutrition package, and the base material of each nutrition package is lactose.
The specific preparation process of the infant formula containing the probiotics and prebiotics composition is as follows:
1) Rough filtration of cow milk: the cow milk is subjected to coarse filtration and degassing by a balance cylinder, is preheated by a plate heat exchanger, and is separated from impurities by a separator.
2) Homogenizing and sterilizing cow milk: part of raw milk after removing impurities enters a homogenizer for homogenization and the other part is not homogenized, and the raw milk and the homogenized raw milk are mixed and enter a sterilizing system for sterilization.
3) Powder adding: the powder raw materials are metered according to the formula, then are added into a powder mixing tank in a unified way through an air conveying system, and are sucked into a vacuum mixing tank through a vacuum system;
4) Melting and oil preparing: placing the grease specified in the formula into an oil melting room according to the formula requirement, keeping the temperature of the oil melting room at 50-90 ℃, after the oil is melted, pumping the oil into a mixed oil storage tank, and pumping the mixed oil into the mixed oil tank through an oil pump according to the formula requirement;
5) Nutrient dissolution and addition: and respectively dissolving the nutrient packages such as calcium powder, vitamins, minerals and the like with purified water, and sequentially adding the dissolved nutrient packages into a mixing tank to obtain mixed feed liquid.
6) Breast milk oligosaccharide dissolution addition: and (3) dissolving part of the mixed liquor in the step (5) for the breast milk oligosaccharide raw material, and then adding the dissolved mixed liquor into a mixing tank to obtain the mixed liquor containing 2' FL.
7) And (3) filtering: filtering the mixed feed liquid containing breast milk oligosaccharide by a filter screen to remove physical impurities possibly brought in the raw materials.
8) Homogenizing: homogenizing the mixed feed liquid by a homogenizer, mechanically treating the fat globules, and dispersing the fat globules into uniform fat globules.
9) Cooling and storing: the homogenized feed liquid enters a plate heat exchanger for cooling: cooling to below 20 ℃, temporarily storing in a pre-storing cylinder, entering the next working procedure within 6 hours, and starting the stirrer according to the set requirement.
10 Concentration sterilization: during production, double-effect concentration is used, the sterilization temperature is more than or equal to 83 ℃, and the sterilization time is 25 seconds. The discharge concentrations were 50% dry matter.
11 Concentrated milk storage, pre-heating filtration, spray drying: the concentrated milk is temporarily stored in a concentrated milk balance tank. Preheating to 60deg.C by scraper preheater, filtering with 1mm pore size filter, spray drying by high pressure pump, and agglomerating fine powder on top of tower or fluidized bed. Air inlet temperature: the temperature of exhaust is at 180 ℃ and 86 ℃, the pressure of the high-pressure pump is 200bar, and the negative pressure of the tower is about-4 mbar.
12 Fluidized bed drying and cooling: and (3) the powder from the drying tower is subjected to fluidized bed (primary) secondary drying and then is cooled to 30 ℃ through a fluidized bed (secondary), so that the milk powder main material is obtained.
13 Split charging: and weighing DHA, ARA or bifidobacterium longum subspecies YLGB-1496 or bifidobacterium lactis according to the formula requirement, sealing and packaging.
14 Dry blending): mixing the weighed DHA, ARA or bifidobacterium longum subspecies YLGB-1496 with the milk powder main material in a dry mixer.
15 Screening powder: the granularity of the milk powder is uniform through the vibrating screen, and the powder slag is scrapped.
16 Powder discharge: and (3) receiving powder by using a sterilized powder collecting box, and conveying the powder from a powder outlet room to a powder feeding room.
17 Powder) is added: and pouring the milk powder into a powder storage tank on a size packaging machine according to the packaging requirement.
18 Packaging: 400 g of automatic packaging machine fills nitrogen for packaging. The oxygen content is lower than 1% when nitrogen is filled. 900 g of iron can is automatically filled with nitrogen and packaged, and the oxygen content is lower than 5%.
19 Boxing: packaging the packaged small bags into a paper box, adding a powder spoon at the same time, and sealing by a box sealing machine.
20 Inspection of the finished product: sampling and checking the packaged product according to a checking plan.
21 Warehousing and storing: the qualified products are stored in warehouse, and the storage is required at normal temperature, and the humidity is less than or equal to 65%.
In the product, the content of the breast milk oligosaccharide 2' FL is about 0.2g/100g; the content of bifidobacterium longum subspecies YLGB-1496 is 1 multiplied by 10 6 CFU/g。
Example 2, infant formula containing probiotic and prebiotic composition (1000 kg prepared):
1000 kg raw milk, 250 kg skim milk powder, 150 kg lactose, 34% whey protein powder WPC 50 kg, 90 kg desalted whey powder D225 kg OPO structural fat 106 kg soybean oil 37 kg corn oil 30 kg alpha-whey protein powder 10 kg beta-casein powder 10 kg fructo-oligosaccharide powder 5kg galacto-oligosaccharide slurry 15 kg soya lecithin 4 kg compounded nutrient 11 kg DHA12 kg ARA 14 kg breast milk oligosaccharide (2' FL) 8kg bifidobacterium longum subspecies YLGB-1496.2 kg and nucleotide 0.65 kg.
Wherein the compound nutrients comprise about 1.5 kg of compound vitamin nutrition package, about 0.75 kg of choline chloride nutrition package, about 5kg of calcium powder nutrition package, about 1 kg of mineral nutrition package, about 0.75 kg of magnesium chloride nutrition package and about 2 kg of potassium chloride nutrition package, and the base material of each nutrition package is lactose. The product preparation process is as in example 1.
In the product, the content of the breast milk oligosaccharide 2' FL is about 0.8g/100g; the content of bifidobacterium longum subspecies YLGB-1496 is 2 multiplied by 10 6 CFU/g。
Example 3 infant formula containing probiotic and prebiotic composition (1000 kg prepared):
1000 kg raw milk, 250 kg skim milk powder, 140 kg lactose, 34% whey protein powder WPC 50 kg, 90 kg desalted whey powder D225 kg OPO structural fat 106 kg soybean oil 37 kg corn oil 30 kg alpha-whey protein powder 10 kg beta-casein powder 10 kg fructo-oligosaccharide powder 5kg galacto-oligosaccharide slurry 15 kg galacto-oligosaccharide slurry 30 kg breast milk oligosaccharide (2' FL) 30 kg compounded nutrient 11 kg DHA12 kg ARA 14 kg bifidobacterium subspecies YLGB-1496.4 kg nucleotide 0.65 kg.
Wherein the compound nutrients comprise about 1.5 kg of compound vitamin nutrition package, about 0.75 kg of choline chloride nutrition package, about 5kg of calcium powder nutrition package, about 1 kg of mineral nutrition package, about 0.75 kg of magnesium chloride nutrition package and about 2 kg of potassium chloride nutrition package, and the base material of each nutrition package is lactose. The product preparation process is as in example 1.
In the product, the content of the breast milk oligosaccharide 2' FL is about 3g/100g; the content of bifidobacterium longum subspecies YLGB-1496 is 4 multiplied by 10 6 CFU/g。
Example 4, infant formula containing probiotic and prebiotic composition (preparation 1000 kg):
1000 kg raw milk, 250 kg skim milk powder, 165 kg lactose, 34% whey protein powder WPC 50 kg, 90 kg desalted whey powder D225 kg OPO structural fat 106 kg soybean oil 37 kg corn oil 30 kg alpha-whey protein powder 10 kg beta-casein powder 10 kg fructo-oligosaccharide powder 5kg galacto-oligosaccharide slurry 15 kg galacto-oligosaccharide 5kg (2'FL 2.65kg,LNT 0.8kg), 11 kg compound nutrient, 12 kg DHA, 14 kg ARA, YGB-1496.4 kg bifidobacterium longum subspecies and 0.65 kg nucleotide.
Wherein the compound nutrients comprise about 1.5 kg of compound vitamin nutrition package, about 0.75 kg of choline chloride nutrition package, about 5kg of calcium powder nutrition package, about 1 kg of mineral nutrition package, about 0.75 kg of magnesium chloride nutrition package and about 2 kg of potassium chloride nutrition package, and the base material of each nutrition package is lactose. The product preparation process is as in example 1.
In the present product, the milk oligosaccharide composition has a 2' FL content of about 0.265g/100g and an LNT content of about 0.8g/100g; the content of bifidobacterium longum subspecies YLGB-1496 is 4 multiplied by 10 6 CFU/g。
Bifidobacterium longum infant subspecies and breast milk oligosaccharide composition efficacy experiments
Probiotic bifidobacterium longum infant subspecies YLGB-1496 the probiotic bacteria tested were cultivated in MRS medium supplemented with 1% cysteine and incubated overnight in an anaerobic environment at 37 ℃. Harvesting thallus, washing with physiological saline solution, adjusting probiotic concentration, inoculating into plates containing nematode growth medium, wherein probiotic concentration is 1×10 in each plate 8 CFU or 1X 10 7 CFU。
The HMO uses distilled water to prepare a breast milk oligosaccharide solution.
Nematode infection model
Obtaining nematode of uniform age, culturing in culture dish containing nematode agar medium (NGM medium containing Escherichia coli OP50 as food), adding different doses of HMO (15 mg/mL) and probiotics (1×10) 8 CFU、1×10 7 CFU) the composition was co-cultured (total volume of liquid added per nematode dish was 10mL. For the HMO test group at a concentration of 15mg/mL, the final amount of HMO in each dish was 150mg. Thus, the ratio of the probiotic to HMO added in each petri dish plate was: 1X 10 8 CFU:150mg, or 1X 10 7 CFU:150 mg). After adult life of the nematodes, they were transferred to dishes inoculated with Staphylococcus aureus ATCC25923, added in an amount of 10 8 ～10 9 CFU/mL to simulate infection with staphylococcus aureus. The case of the intervention (probiotics and/or HMO) added during the infection phase for each group was the same as for the culture phase, respectively. Two controls were used, each in the absence of pathogenic bacteria (E.coli OP50 in the nematode dishes) and in the presence of pathogenic bacteria, staphylococcus aureus infection, but without any intervening material (dishes containing Staphylococcus aureus only). Addition of culture stageThe group of the intervention without any intervention at the infection stage, and the group without intervention at the incubation stage with intervention at the infection stage.
After several days of incubation, nematodes were counted daily for their viability. Nematodes are considered dead if they do not react to platinum wires. Two independent determinations were made for each condition.
Statistical comparative analysis of survival curves was performed and log rank survival significance analysis was performed using GraphPad Prism 4 statistical software package. Significant differences in survival on the third day of infection were analyzed with One-way ANOVA. The survival data for each group on each day was statistically analyzed with Two-way ANOVA and significantly analyzed with Dunnett' posthoc test.
Experimental results
As shown in FIG. 1, 15mg/mL HMO 2' -FL and YLGB-1496 (10 8 CFU) has remarkable protection and promotion effects on survival rate of nematodes infected by staphylococcus aureus (P)<0.0001 And the protection effect of the two is relatively similar. And 10 (10) 7 The YLGB-1496 strain also has better protection effect on the nematode survival rate in the condition, although the effect is obviously lower than 10 8 Survival under conditions (P<0.05). When 2' -FL and YLGB-1496 Strain (10 8 CFU) to form a composition, in which case the effect on the increase in survival of nematodes when infected with staphylococcus aureus is significantly better than either of these two substances alone, i.e. the strain alone or HMO alone, and the composition achieves a 22% increase in survival compared to the starting material alone (P<0.001). It can be seen that when 2' -FL and YLGB-GB1496 form a composition, the two show good synergistic effect.

Claims (5)

1. Use of a combination of Bifidobacterium longum subspecies infantis and breast milk oligosaccharides for the preparation of a formula having an effect of improving the resistance of organisms to Staphylococcus aureus infection, wherein 1X 10 Bifidobacterium longum subspecies infantis is contained per 100g of formula 6 CFU~1×10 12 CFU，0.1 g-3 g of breast milk oligosaccharide;
wherein the bifidobacterium longum subspecies infancy is the bifidobacterium subspecies infancy with the preservation number of CGMCC No.21109Bifidobacterium longum subsp. infantis) YLGB-1496 strain;
the breast milk oligosaccharide is 2' -fucosyllactose.
2. The use according to claim 1, wherein the total protein content in the formula milk powder is 10-20 g/100g, and the whey protein accounts for 40-70% of the total protein; the fat content is 15 g-30 g/100g; the carbohydrate content is 50 g-70 g/100g.
3. The use according to claim 1, wherein the formula milk powder has a linoleic acid content of 2700-4500 mg/100g and an α -linolenic acid content of 270-450 mg/100g.
4. The use according to claim 1, wherein the formula is an infant formula or a non-infant formula.
5. The use according to claim 1, wherein the formula further comprises one or more of nutrients, dietary fiber, inositol, taurine, l-carnitine, docosahexaenoic acid, eicosatetraenoic acid, lutein, nucleotides, lactoferrin.
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